A new method for the adaptive determination of optimum pH and temperature

An adaptive control algorithm for the on-line determination of optimal temperature or pH for biomass production in a continuous fermentor is presented. The algorithm requires no prior information and uses a dynamic Hammerstein model to identify parameters and to estimate an optimal steady-state control value. A check of the estimated performance measure second derivative is included to ensure that the target extremum is an optimum. The process is driven towards this optimum with a variable step size that depends on the quality of the on-line identified model. Numerical simulations are performed on a dynamic chemostat model that incorporates a metabolic time delay. The algorithm successfully finds the optimum temperature or pH values and maintains the reactor at the optimum steady state.     

A common source of error in pH measurements.

Glass-electrode assemblies in which the reference half-cell contains a porous ceramic type of liquid junction are likely to produce misleading pH measurements under normal service conditions. The error arises from substantial liquid-junction potentials, associated with the porous ceramic plug, which vary with the ionic composition of the solution under test. The error is not revealed by conventional two-point calibration procedures, since the majority of standard buffer solutions have a similar total ionic strength, but will nevertheless be present when the unknown solution differs in ionic strength from the standardizing buffers. The size of the error is proportional to the ratio between the salt concentration in the standard buffers and the concentration present in the unknown solution, and varies from one electrode specimen to another. The fault was present in 24 out of 30 electrodes in normal use selected at random from seven laboratories, and the mean error was 0.2pH unit per 10-fold salt-concentration difference between standard and test solutions. It is estimated that errors of this order must be widespread in the recent literature. Older pH determinations are likely to be more reliable, since the original reference electrode design with a free-flowing liquid junction is apparently free from the artefact.     

A new method for temperature compensation of electrical conductivity using temperature-fold dependency of fresh water

A new method for temperature compensation of electrical conductivity (EC) is proposed. A relationship between temperature and EC was investigated in detail by a simple experiment using natural and artificial fresh water. Results showed that the temperature dependency of EC varied from 0.03 to 0.02, with temperature increasing from 3°C to 35°C. This means that a traditional method for compensating for EC using a constant coefficient is invalid, so that a temperature-fold dependency must be taken into consideration to standardize EC to a common temperature.     

A mathematical model for pH patterns in the rhizospheres of growth zones

In the classical model by Nye (1981), the main process for the change in pH across the rhizosphere is assumed to be diffusion. The classical model focuses on the non-growing part of the root and assumes that the distribution of ion fluxes along the root is spatially uniform. We consider the rhizosphere of the growth zone and take into account the root growth rate and spatially varying flux along the root surface. We present both analytical (dimensional analysis) and experimental (computational) evidence of the importance of taking into account the root growth rate. We describe a conceptual and mathematical model to analyse the pH field around the root tip over time. The model is used with published data to show that, for typical growth rates in sandy soil, the pH field becomes steady (independent of time) after 6 h. Dimensional analysis reveals that a version of the Péclet number, related to the quotient of root elongation rate and proton diffusivity, can be used to predict the extent of the rhizosphere and the time required for it to become steady. For Péclet numbers much greater than 1 (soils), the root influences soil pH for distances on the millimetre scale. In contrast, for Péclet numbers much less than one (agar, aqueous solution), the root influences substrate pH for radial distances on the scale of centimetres. We also present some evidence that agar-contact techniques to measure the soil pH may not be appropriate for measuring the millimetre-scale gradients in soil pH.     

Effect of ambient temperature and attachment method on surface temperature measurements

Accurate measurement of skin surface temperature is essential in both thermo-physiological and clinical applications. However, a literature review of the last two decades of physiological or clinical research revealed an inconsistency or a lack of information on how temperature sensors were attached to the skin surface. The purpose of this study was to systematically compare and quantify the performance of different commercially available temperature sensors and their typical attachment methods, and, secondly, to provide a time-efficient and reliable method for testing any sensor-tape combination. In conclusion, both the sensor type and the attachment method influenced the results of temperature measurements (both its absolute and relative dimensions). The sensor shape and the contact of its sensing area to the surface, as well as the conductance of the tape were the most important parameters to minimise the influence of environmental conditions on surface temperature measurement. These results suggest that temperature sensors and attachment methods for human subject and manikin trials should be selected carefully, with a systematic evaluation of the sensor-tape system under conditions of use, and emphasise the need to report these parameters in publications.     

A mathematical method for analysing plasmid stability in micro-organisms

A mathematical model describing the instability of plasmids in micro-organisms has been developed. The model is based on the assumption that the overall causes of plasmid instability are described by the segregational instability of the plasmid, R (i.e. the rate at which plasmid-free cells are generated from plasmid-bearing cells), and the growth rate difference, d mu (i.e. the difference in growth rate between plasmid-free and plasmid-bearing cells). A method for determining the values of R and d mu (accompanied by 95% confidence limits) for any plasmid-bearing micro-organism is described. This method is based on the observation that, depending on the plasmid, various exponential patterns of plasmid instability are observed. The stability of Escherichia coli 1B373(pMG169), where d mu much greater than R, and E. coli RV308(pHSG415), where R much greater than d mu, are analysed in order to demonstrate the method.     

A multichannel sensor for temperature measurements in shallow waters

A multichannel temperature sensor is presented; it is suitable for almost simultaneous measurements at various depths and time intervals. It allows automatic registration of smallscale time/depth variations of temperature in floating masses of filamentous algae and similar thermally stratified systems. An example is given by a 4-day record of the temperature regime in a floating mass ofOedogonium species in May.     

A facile HPLC method for optical purity and quantitative measurements of phenylalanine from the hydrolyzed aspartame under different pH and temperature after its derivatization with a fluorescent reagent

Summary. In this paper, the artificial sweetener aspartame is deliberately hydrolyzed under different pH and temperature in the matrix, and time period for the hydrolysis. The HPLC analysis is then performed to quantitatively measure the amount and the optical purity of phenylalanine produced as a result of hydrolysis in the matrix after its functionalization with a fluorescent reagent. The results show that the amount of phenylalanine in the matrix is affected by the pH variation during the hydrolysis and found increased in low pH conditions. High temperature or long time periods for the decomposition also increases the amount, which indicates that beverages and foods containing aspartame as a sweetener may not be safe for phenylketonuria patients to consume if they are stored under these conditions. Conversely, the optical purity of phenylalanine, expressed as the percentage of d-enantiomer, is not affected by pH variations. However, it decreases as the length of time elapsed is increased or surrounding temperature is elevated during the decomposition.     

A stereophotogrammetric method for measurements of ligament structure

A stereophotogrammetric method is presented to reconstruct the course of a curve in the three-dimensional space. This method is exclusively suitable as a non-destructive tool to determine the surface fiberstructure of ligaments, tendons and other organised collagenous structures. In addition, it is a convenient tool to measure the geometry of articular surfaces and other complicated surface shapes.     

pH-sensitive glass microelectrodes and intracellular pH measurements.

1. Some properties of the open-tipped, uninsulated, pH-sensitive glass microelectrode were examined in several electrical experiments. 2. Based on these observations, technical and theoretical problems were considered for application to the pH measurement in small cells. 3. The intracellular pH, (pH)i, of the epithelial cell in rat duodenum measured was approximately 7.0. A reduction in (pH)i was apparent (about 0.3) with the addition of 20 mM-glucose to the bathing fluid. 4. It was concluded that with certain limitations such uninsulated, open-tipped microelectrodes may be successfully utilized for intracellular pH measurements.     

A mathematical model for the immunosenescence.

We propose a model for the dynamics of the immune system by considering the subpopulations of virgin and memory T lymphocytes on a time scale corresponding to the human life span. In the deterministic balance equation we introduce a fluctuating term in order to take into account the chronic antigenic stress. Starting from the hypothesis that the depletion of virgin cells with cytotoxic properties (CD8+) is a mortality marker, the model provides survival curves quite similar to the demographic curves.     

A multicouple probe for temperature gradient measurements in biological materials

An easy-to-make, sensitive, thin, flexible, multisensor probe for in vivo tissue temperature profile measurement is described. It is essentially a multijunction thermocouple (i.e., a multicouple) of type-T composition. Enamel-insulated copper wires (38 gauge) were soldered 5 mm apart to one common uninsulated constantan wire (36 gauge) and introduced into a polyethylene tube sealed at one end. The total outside diameter of the multicouple probe is less than 1 mm, and the maximum number of junctions using the specified wire sizes is approximately 16. This design permits the instantaneous measurement of a tissue temperature profile at 5-mm intervals over a distance of approximately 8 cm. An extensive calibration for the thermal conductivity effect (k effect) along the multicouple wires by means of a limb model is presented. The results show that the temperature readings of the individual junctions are significantly affected by the k effect when a thermal gradient exists along the multicouple, as is usually the case during tissue temperature measurements. However, calibration of the multicouple for the k effect yields a measurement accuracy of +/- 0.1 degree C under a wide range of gradients. This probe can be implanted in tissues to measure thermal gradients under different physiological conditions.     

A rapid method for measuring intracellular pH using BCECF-AM

A rapid intracellular pH (pH(i)) measurement method based on initial rate of increase of fluorescence ratio of 2',7'-bis(2-carboxyethyl)-5,6-carboxyfluorescein upon dye addition to a cell suspension in growth medium is reported. A dye transport model that describes dye concentration and fluorescence values in intracellular and extracellular spaces provides the mathematical basis for the approach. Experimental results of ammonium chloride challenge response of the two suspension cells, Spodoptera frugiperda and Chinese hamster ovary (CHO) cells, successfully compared with results obtained using traditional perfusion method. Since the cell suspension does not require any preparation, measurement of pH(i) can be completed in about 1 min minimizing any potential errors due to dye leakage.     

Studies on on-line bioreactor identification. IV. Utilization of pH measurements for product estimation

Relationships between the total rate of biomass growth and the rate of ammonia addition to a fermentor for pH control are presented. These equations make use of the concept of reaction invariants and provide the additional information needed for bioreactor identification. They are especially useful when the RQ measurement is not sufficient for this purpose, such as when sensitivities arise with the measured values of the respiratory quotient or when fermentation products are formed. The cases of batch, fed-batch and continuous fermentations, forming products with or without acidic/basic properties are considered. The derived relationships were successfully tested with nonbiological acid-base continuous flow reaction systems and subsequently applied to the identification of the continuous yeast fermentation of glucose to ethanol. Results of these experimental studies are also presented.     

Rapid method for measuring intracellular pH in vivo.

Intracellular pH (pHi) was simultaneously measured in 6 normal tissues and a malignant tumour of rats by a rapid triple isotope technique, based on the in vivo distribution of 5,5-dimethyl-2,4-oxazolidinedione-2-14C (DMO), tritiated water and sodium chloride-36. Results compared favourably with pH measured directly in the same rat by capillary glass electrode, and with values of other workers for pHi in rat tissues. Mean pHi of normal tissues was close to pH 7, and in each organ there was a linear relationship between pHi and extracellular pH (pHe) over the normal range of pHe encountered (pH 6.9-7.6). Organ pHi altered in response to administration of NH4Cl or NaHCO3 to the host.